Cells for Therapy of the Heart

Abstract

According to the invention fibroblast-like cells obtained from heart muscle biopsies, which are CD90 negative, CD105 positive, CD117 negative and/or CD166 positive as well as cell preparations of such cells for therapy of heart diseases as well as a method for providing the latter are disclosed. The cells according to the invention are characterized by a good cultivability in cell culture. Furthermore a method for obtaining the cells and cell preparations according to the invention are disclosed.

We claim: 
     
         1 . A method for obtaining a cell preparation comprising isolated fibroblast-like cells, the method comprising:
 a) in a first cell culture step, culturing a tissue sample obtained from mammalian heart muscle tissue under conditions suitable for culture of mammalian cells in cell culture medium, in a cell culture container having a solid surface;   b) detaching cells that have grown out of the tissue sample and that adhere to the solid surface in a passage step by means of limited proteolysis, isolating the detached cells;   c) culturing the sorted cells in a second cell culture step;   thereby obtaining the cell preparation that is CD90 negative; CD105 positive; CD106 negative and CD117 negative.   
     
     
         2 . The method according to  claim 1 , wherein said fibroblast-like cells are characterized by
 a) an expression of one or more genes selected from ACTC1, POP3, VE-Cadherin (CDH5) and CDH13 and/or   b) a lack of expression of NPNT and/or TERT.   
     
     
         3 . The method according to  claim 2 , wherein said fibroblast-like cells are characterized by expression of ACTC1, POP3, VE-Cadherin (CDH5) and CDH13 and by lack of expression of NPNT and TERT. 
     
     
         4 . The method according to  claim 1 , wherein said tissue sample comprises or essentially consists of endomyocardial cells. 
     
     
         5 . The method according to  claim 1 , wherein said tissue sample comprises or essentially consists of atrial appendage cells, and subsequent to said passage step, CD90 positive cells are excluded from the detached cells via magnetic activated cell sorting or fluorescence activated cell sorting in a CD90-sorting step. 
     
     
         6 . The method of  claim 1 , wherein the tissue sample is digested, prior to the first cell culture step, by one or more connective tissue digesting enzymes 
     
     
         7 . The method of  claim 6 , wherein the connective tissue digesting enzyme comprises at least one of trypsin-EDTA and collagenase IV, and wherein the time limited digestion is less than 10 minutes at an activity of 0.05 to 0.25 u/500 ml for trypsin and/or 0.2 to 4.5 u/ml for collagenase IV. 
     
     
         8 . The method of  claim 1 , wherein the cell culture medium of either the first cell culture step, the passage step or of both steps does not contain any of cardiotrophin, thrombin and mercaptoethanol. 
     
     
         9 . The method of  claim 1 , wherein the first cell culture step (a) has a duration of 7 to 15 days. 
     
     
         10 . The method of  claim 1 , wherein the cells are maintained in said second cell culture step for a period of 7 to 15 days or 7 to 12 days. 
     
     
         11 . The method of  claim 1 , wherein the passage step (b) is conducted once the cells adhering to the solid surface are at least 70% confluent. 
     
     
         12 . A cell preparation produced by the method according to  claim 1 . 
     
     
         13 . The cell preparation according to  claim 12 , wherein the cells contained in the cell preparation are more than 90% CD90 negative, more than 90% CD105 positive, and more than 50% CD117 negative. 
     
     
         14 . The cell preparation according to  claim 12 , wherein the cells contained in the cell preparation are more than 95% CD90 negative, more than 95% CD105 positive, more than 60% CD117 negative, and more than 50% CD166 positive. 
     
     
         15 . The cell preparation according to  claim 12 , wherein the cells contained in the cell preparation are more than 90% CD34 negative and more than 90% CD45 negative. 
     
     
         16 . The cell preparation according to  claim 12 , wherein the cells contained in the cell preparation are more than 95% CD90 negative, and the CD90 negative portion of the cell preparation is more than 90% CD105 positive. 
     
     
         17 . The cell preparation according to  claim 12 , wherein the cells contained in the cell preparation are more than 95% CD90 negative, and the CD90 negative portion of the cell preparation is more than 90% CD105 positive and more than 50% CD117 negative. 
     
     
         18 . The cell preparation according to  claim 12 , wherein the cells contained in the cell preparation are more than 98% CD90 negative, and the CD90 negative portion of the cell preparation is more than 95% CD105 positive, more than 60% CD117 negative and more than 50% CD166 positive. 
     
     
         19 . The cell preparation according to  claim 12 , wherein the cells contained in the cell preparation are more than 95% CD90 negative, the CD90 negative portion of the cell population is more than 90% CD105 positive and the portion of the cell preparation being CD90 negative and CD105 positive at the same time is more than 60% CD117 negative. 
     
     
         20 . The cell preparation according to  claim 12 , wherein the cells contained in the cell preparation are more than 90% negative for CD106 (VCAM1) and/or the cells contained in the cell preparation express ACTC1 and/or the cells contained in the cell preparation express POP3 and/or the cells contained in the cell preparation express POP3 and/or the cells contained in the cell preparation express VE-Cadherin (CDH5) and/or the cells contained in the cell preparation express CDH13 and/or the cells contained in the cell preparation do not express NPNT and/or the cells contained in the cell preparation do not express TERT. 
     
     
         21 . A method for treatment of cardiac disease, comprising
 administering to a subject in need thereof a therapeutically effective amount of a cell preparation comprising isolated fibroblast-like cells, obtained from mammalian heart muscle tissue, wherein the isolated fibroblast-like cells are CD90 negative; CD105 positive; CD106 negative and CD117 negative.